羊耳菊提取物主要活性成分在大鼠体内的组织分布研究

为了研究大鼠灌胃羊耳菊提取物后7个指标成分在体内的组织分布情况,实验建立同时测定大鼠组织中4,5-二咖啡酰基奎宁酸、新绿原酸、绿原酸、3,4-二咖啡酰基奎宁酸、1,3-二咖啡酰基奎宁酸、隐绿原酸、木犀草苷的UPLC-MS/MS方法,将羊耳菊提取物灌胃给予SD大鼠,分别于给药0. 5、1. 5、5 h取其主要脏器和组织,采用UPLC-MS/MS测定各时间点下7个指标成分在脏器和组织中的分布情况。大鼠灌胃羊耳菊提取物后,对于新绿原酸,其浓度0. 5 h在小肠、肾、肺、肝达到峰值; 1. 5 h在胃、肌、脾达到峰值; 5 h在心达到峰值。对于绿原酸,其浓度0. 5 h在小肠、肾、肺、心达到峰值; 1. 5 h在胃、肌、脾、肝达到峰值。对于隐绿原酸,其浓度0. 5 h在小肠、肾、肺达到峰值; 1. 5 h时在心、肝、脾、肺、胃达到峰值。对于1,3-二咖啡酰基奎宁酸,其浓度0. 5 h在心、肺、肾、小肠达到峰值; 1. 5 h在肝、脾、肌、胃达到峰值。对于3,4-二咖啡酰基奎宁酸,其浓度0. 5 h在小肠和肾达到峰值; 1. 5 h在肝、脾、肌、胃达到峰值; 5 h在心、肺达到峰值。对于4,5-二咖啡酰基奎宁酸,其浓度0. 5 h在小肠、肾、心达到峰值; 1. 5 h在肝、脾、肌、胃达到峰值; 5 h在肺达到峰值。对于木犀草苷,其浓度0. 5h在小肠和心达到峰值; 1. 5 h在肝、脾、胃达到峰值; 5 h在肺和肾达到峰值。7个指标成分可迅速、广泛地分布在各组织器官中,脑组织中未检测到该7种成分。7种成分主要分布在胃、小肠和肾组织中,对肾脏表现出较强的亲和力,推测肾脏可能是羊耳菊的主要排泄器官之一。     

牡丹籽饼粕中芍药苷类成分及其大孔吸附树脂纯化研究

采用乙醇提取,树脂纯化,HPLC制备以及LC-MS和1H NMR鉴定,从牡丹籽粕的醇提物中分离纯化了4种主要成分,分别为6'-O-β-D-葡萄糖芍药内酯苷、芍药内酯苷、β-gentiobiosylpaeoniflorin和芍药苷。对大孔吸附树脂法纯化芍药苷类成分的条件进行了试验,从4类11种树脂中筛选出HPD-200A型大孔吸附树脂,其较优的吸附分离条件为:上样液浓度(芍药苷)8.0 mg/mL,上样体积为4.5倍床体积(BV),流速为1/16 BV/min,洗脱剂乙醇溶液浓度为50%(v/v),洗脱体积为4 BV,流速为1/16 BV/min。此条件下所得提取物中含芍药苷32.3%、芍药内酯苷16.5%、6'-O-β-D-葡萄糖芍药内酯苷8.02%、β-gentiobiosylpaeoniflorin 6.63%。     

洛伐他汀在巴马香猪体内的分布和排泄

目的研究洛伐他汀在巴马香猪体内的分布和排泄特点。方法以抗动脉粥样硬化药物洛伐他汀为模型药,选择健康6月龄雄性巴马香猪为实验对象,经灌胃途径给药(45 mg/kg或2.4 mg/kg),采用RP-HPLC方法测定各组织及体液中的药物浓度,并对其分布和排泄过程进行研究。血浆蛋白结合率通过透析法测定。结果给药后,洛伐他汀快速分布到贲门、胃、小肠、肝、大肠、胰、前列腺、肺、肾、心、肌肉、睾丸、肾上腺、膀胱、脑和脾。以胃、肠、肝组织中药物浓度较高。单次给药4h后,贲门、胃、小肠、肝、心、肾上腺、膀胱药物浓度同给药后1h相比略有下降,其余组织均高于1 h。血浆蛋白结合率为95%以上,同正常人血浆非常一致。96 h尿中累积排泄量为给药量的7.4%,原形药经胆汁及粪排泄量达到80%以上。结论洛伐他汀在巴马香猪体内同人的分布排泄和血浆蛋白结合率相似,均在组织中广泛分布,血浆结合率达到95%以上,主要经胆汁和粪排泄。     

不同日龄大白猪视黄酸受体α基因组织表达

研究采用RT-PCR方法对大白猪的视黄酸受体α基因在1日龄、90日龄、180日龄、270日龄和360日龄的心、肝、胃、脾、肾、肺、大肠、小肠、肌肉、子宫、卵巢共11个组织的表达情况进行了研究。结果表明,RARαmRNA在肝、脾、肾、大肠、小肠、子宫和卵巢中持续表达,其中脾、大肠和小肠是持续高表达;180日龄时,所有组织的RARαmRNA的表达量普遍降低;360日龄时,所检的11个组织均高水平表达该基因。     

黄芪甲苷在正常小鼠与2型糖尿病肾病 db/db小鼠体内分布差异的比较研究

为研究黄芪甲苷(Astragaloside IV,AS-Ⅳ)在正常小鼠(db/m)和2型糖尿病肾病(db/db)小鼠体内的组织分布差异,为AS-Ⅳ抗2型糖尿病肾病的临床运用及新药研发提供实验依据。以尾静脉注射给予2型糖尿病肾病(db/db)和正常小鼠(db/m)小鼠AS-Ⅳ,剂量为8 mg/kg。于15 min、2 h、4 h时处死小鼠,收集心、肝、脾、肺、肾、胃、小肠、脑、肌肉组织,采用HPLC-MS/MS法测定各组织中AS-Ⅳ含量,对比AS-Ⅳ在正常及病理状态下,在各组织中的分布差异。在2型糖尿病肾病状态下,AS-Ⅳ在肝、脾、肺、肾组织中浓度分别为349.72±70.72、370.69±45.46、5 830.65±581.75、4 290.63±485.34 ng/mL;正常状态下,AS-Ⅳ在肝、脾、肺、肾组织中浓度分别为202.47±47.96、267.92±41.24、4 725.80±867.51、2 354.55±256.11 ng/mL,在2型糖尿病肾病状态下,AS-Ⅳ在肝、脾、肺、肾组织中浓度显著高于其在正常组织中浓度(P0.05)。静脉注射给药后,肺、肾、心、胃、脾是其主要分布器官,其中在肺、肾组织中浓度最高;病理状态下,AS-Ⅳ组织分布发生了一定改变,为AS-Ⅳ防治2型糖尿病肾病的临床合理应用及开发提供实验依据。     

基于HPLC指纹图谱的亳白芍不同炮制品标准汤剂7个成分含量测定

建立亳白芍不同炮制品标准汤剂HPLC指纹图谱,其中生白芍、炒白芍和酒白芍标准汤剂分别标定16、14和13个共有峰,同时测定亳白芍不同炮制品标准汤剂中7种化学成分(没食子酸、氧化芍药苷、芍药内酯苷、芍药苷、苯甲酸、1,2,3,4,6-五没食子酰葡萄糖和苯甲酰芍药苷)含量,经炒制后标准汤剂中芍药内酯苷和苯甲酸含量升高,氧化芍药苷、芍药苷含量降低,酒白芍标准汤剂中芍药苷含量升高,苯甲酸含量降低,并对含量测定结果进行聚类分析,同一批亳白芍及其炮制品可聚为一类,但聚类距离缩小后生品和炮制品各聚为一类。建立的HPLC指纹图谱和含量测定方法具有良好的重现性,且简便快速,二者结合可直观反映出亳白芍炒制、酒制后的变化差异。     

相同基源的赤芍和白芍中芍药苷含量的比较

通过采用HPLC法比较了中江产芍药的根部不同加工品-赤芍和白芍中芍药苷的含量,以期为研究二者功效差异的具体原因奠定基础.结果表明,赤芍和白芍的化学成份及其含量均存在一定差异,大、中、小三个规格的赤芍和白芍中芍药苷含量分别2.97%、2.94%、2.98%和1.86%、1.82%、1.91%;白芍中芍药苷含量低于赤芍,芍药苷含量与规格无明显关系,化学成份及其含量的差异可能是二者功效差异的具体原因之一.     

近红外光谱法快速测定白芍中芍药苷含量

本研究旨在应用近红外光谱法建立一种白芍药材中芍药苷含量的快速测定方法。利用HPLC测定样品中芍药苷含量,并以其作为参考值,运用偏最小二乘法(PLS)建立芍药苷含量与近红外光谱之间的多元校正模型,对未知样品进行含量预测。结果表明,所建芍药苷定量分析模型的相关系数(R2)、内部交叉验证均方差(RMSECV)、校正均方差(RMSEC)分别为0.99395、0.33068、0.0563;经内部验证,模型的预测均方差(RMSEP)和平均回收率分别为0.0756和100.07%。该方法操作简便,无污染,结果准确可靠,可用于白芍中芍药苷含量的快速测定。     

自组装海藻酸钠纳米粒在正常小鼠体内分布的研究

目的:研究自组装海藻酸纳米粒(sSAN)在小鼠的体内分布情况,探讨sSAN作为维生素D3药物载体的可行性。方法:用异硫氰基荧光素(FITC)标记sSAN和负载维生素D3的海藻酸纳米粒(sSAN-VD3),将两种标记好的纳米粒分别给予小鼠灌胃,在不同的时间将小鼠处死,分别取血清和肝、肺、肾、脾,各脏器经匀浆后,用荧光分光光度计测定其荧光强度,计算血清和各组织中sSAN和sSAN-VD3的含量。结果:经灌胃给药后在小鼠血清、肝、肾、肺中均检测到上述药物,而脾中没有检测到。给药后0.5h和1h,sSAN-VD3-FITC及sSAN-FITC在肝、肺和血清中的含量持续增加,以1h时达峰值浓度,给药2h、4h后,两种制剂的浓度逐渐降低。在肾脏中的含量随着时间的延长逐渐增加,于2h时达峰值浓度,随后逐渐降低。结论:sSAN及sSAN-VD3经小鼠灌胃给药后均可吸收入血,而且口服吸收后在肝、肺、肾和血清中均有一定的分布。     

芍药苷对结肠癌SW480细胞增殖、侵袭、迁移的影响

目的观察芍药苷对人结肠癌SW480细胞株增殖、侵袭、迁移的影响,探究其干预机制。方法含10%胎牛血清的DMEM/F12培养基常规培养人结肠癌SW480细胞株,CCK-8以及EdU-488法检测芍药苷对SW480细胞增殖的影响,Transwell小室检测芍药苷对SW480细胞侵袭、迁移的影响,Westernblot法检测beclin1、Bcl-2蛋白的表达。结果不同浓度芍药苷分别处理24h、48h、72h的结肠癌SW480细胞增殖活性受到显著抑制:相比较对照组,160μg/ml芍药苷处理48h后,SW480细胞内黄绿色荧光减弱,细胞增殖率显著下降,为(58.91±4.99)%;SW480细胞的侵袭细胞数、迁移细胞数显著下降:侵袭抑制率为26.50%,迁移抑制率为24.67%;beclin1蛋白表达高于对照组,Bcl-2蛋白表达低于对照组,beclin1与Bcl-2蛋白表达呈负相关。结论芍药苷能够抑制结肠癌SW480细胞增殖、侵袭和迁移,其机制可能通过抑制Bcl-2蛋白表达,上调beclin1蛋白的表达。     

On the metabolism of amygdalin. 2. The distribution of beta-glucosidase activity and orally administered amygdalin in rats

The organs of 15-day-old rats had the highest capability to hydrolyze amygdalin and prunasin, and most of this activity is concentrated in the tissues of the small and large intestines. The activity decreased with age. In adult rats, the ability of the organs to hydrolyze prunasin is higher than that of amygdalin and is concentrated in the spleen, large intestine, and kidney (35.0, 15.0, and 8.9 micrograms prunasin hydrolyzed . h-1 . g tissue-1). Minced tissues of the liver, spleen, kidney, and stomach contain more hydrolytic capability than the homogenate of these organs, while the reverse is the case with the small and large intestines. When 30 mg amygdalin was orally administered to adult rats, its distribution after the 1st h was as follows: stomach (0.89 mg), small intestine (0.78 mg), spleen (0.36 mg), large intestine (0.30 mg), kidney (0.19 mg), liver (0.10 mg), and serum (5.6 micrograms/mL). At the end of the 2nd h, the highest amygdalin content was found in the large intestine (0.79 mg).     

In vitro metabolism of opioid tetrapeptide agonists in various tissues and subcellular fractions from rats

The metabolism of three mu-selective opioid tetrapeptide agonists, Tyr-D-Arg-Phe-Nva-NH(2) (TArPN), Tyr-D-Arg-Phe-Phe-NH(2) (TArPP), and Tyr-D-Ala-Phe-Phe-NH(2) (TAPP), was investigated in different rat tissues. High metabolic activity (<20% peptide remaining after 30 min) was found against the three peptides in the kidney homogenate and against TArPN in spleen homogenate. Low metabolic activity (>80% peptide remaining after 30 min) was found for all peptides in brain homogenate and plasma, and for TArPN and TArPP in blood. The other tissue homogenates, prepared from the small and large intestine, liver and lung, all exhibited intermediate metabolic activity (20-80% peptide remaining after 30 min) against the peptides. In all tissues investigated, the tetrapeptides were metabolized at the C-terminal amide by deamidation.A further in depth metabolic investigation was performed in subcellular fractions isolated from three tissues (small intestine, liver and kidney). In the liver, the deamidation was predominantly localized to the mitochondrial/lysosomal fraction, while hydrolysis at the N-terminal Tyr residue was the major metabolic pathway in the microsomal/brush-border membrane fraction from the kidney and small intestine.     

Assessment of genomic imprinting of PPP1R9A, NAP1L5 and PEG3 in pigs

Imprinted genes play significant roles in the regulation of fetal growth and development, function of the placenta, and maternal nurturing behaviour in mammals. At present, few imprinted genes have been reported in pigs compared to human and mouse. In order to increase understanding of imprinted genes in swine, a polymorphism-based approach was used to assess the imprinting status of three porcine genes in 12 tissue types, obtained from F1 pigs of reciprocal crosses between Rongchang and Landrace pure breeds. In contrast to human and mouse homologues, porcine PPP1R9A was not imprinted, and was found to be expressed in all tissues examined. The expression of porcine NAP1L5 was detected in pituitary, liver, spleen, lung, kidney, stomach, small intestine, skeletal muscle, fat, ovary, and uterus, but undetectable in heart. Furthermore, porcine NAP1L5 was paternally expressed in the tissues where it's expression was observed. For PEG3, pigs expressed the paternal allele in skeletal muscle, liver, spleen, kidney, and uterus, but biallele in heart, lung, fat, stomach, small intestine, and ovary. Our data indicate that tissue distribution of the three gene differs among mammals, and the imprinting of NAP1L5 and PEG3 is well conserved.     

Ethanol and the antioxidant defense in the gastrointestinal tract.

Ethanol is known to have profound actions on the gastrointestinal tract. The present study was undertaken to examine the effects of ethanol on some of the natural antioxidant defensive enzymes in the gastrointestinal tract; the activities of these enzymes in the liver and the brain were also measured for comparison with those in the gastrointestinal tract. Oral administration of absolute ethanol induced severe gastric mucosal lesions and also damage in the small intestine, however the total superoxide dismutase was unaffected in the tissues measured. The glucose-6-phosphate dehydrogenase activity was reduced only in the stomach while the total glutathione was elevated in the small intestinal mucosa. The catalase activities were activated in the stomach, small and large intestines, and brain, but not in the liver which contained the highest concentration of the enzyme. The present findings indicate that endogenous hydrogen peroxide may be an important damaging agent towards biomolecules in different organs and the removal of this by catalase represents an important defensive mechanism against ethanol toxicity.     

半滑舌鳎miR-200a和miR-200b前体克隆及其免疫应答分析

为了探究半滑舌鳎（Cynoglossus semilaevis）miR-200a和miR-200b在免疫应答中的作用,采用PCR方法克隆了半滑舌鳎miR-200家族的miR-200a和miR-200b的前体序列,长度分别为82和88 bp;用The mfold Web Server和Clustalx1.83软件对其前体序列进行了二级结构和同源性分析,miR-200a和miR-200b都具有典型的颈环结构,与其他物种具有较高的同源性。qRT-PCR分析结果显示,miR-200a和miR-200b在健康半滑舌鳎13种组织（肝脏、肠、脾脏、头肾、后肾、鳃、血液、脑、皮肤、肌肉、胃、心脏和卵巢）中均有表达,miR-200a在头肾中表达量最高,在血液中表达量最低,miR-200b在肝脏中表达量最高,在肌肉中表达量最低;miR-200a和miR-200b在鳗弧菌（Vibrio anguillarum）感染半滑舌鳎后不同时间点的4种免疫相关组织（肝脏、肠、脾脏和头肾）中的表达呈现出先上调后下降的规律,但表达达到峰值的时间点有所不同。miR-200a在肝脏和脾中的表达峰值出现在鳗弧菌感染后6h,在肠和头肾中则是鳗弧菌感染后12h,miR-200b在肠、脾和头肾中均在鳗弧菌感染后12h达到表达高峰;miR-200a和miR-200b在脂多糖（LPS）、肽聚糖（PGN）、葡聚糖（WGP）、聚肌胞苷酸（poly I:C）4种病原模拟物刺激后的半滑舌鳎肝脏细胞系中呈现出上调表达趋势,其中Poly I:C刺激半滑舌鳎肝脏细胞系后miR-200a上调表达趋势明显,6h的表达量为0h的9倍,在WGP刺激半滑舌鳎肝脏细胞后miR-200b上调表达趋势明显,2h的表达量为0的9倍。研究结果为揭示miRNA在半滑舌鳎免疫应答中的作用提供了科学依据。     

Expression of gonadotrophin-releasing hormone binding sites in somatic tissues of the gilthead seabream (Sparus aurata): a quantitative autoradiographic study

In this study, we have analysed the expression of gonadotrophin-releasing hormone (GnRH) binding sites in somatic tissues (intestine, liver, gill, skeletal muscle, ovary, heart, stomach, kidney and spleen) of the gilthead seabream, Sparus aurata using 3-[125I]iodototyrosyl5-mammalian GnRH and auto-radiographic techniques. The qualitative and quantitative analysis showed the existence of a basal expression of specific GnRH binding sites in intestine, skeletal muscle, ovary, stomach and spleen. Furthermore, our data suggest that the level of expression of GnRH binding sites can be significantly enhanced by GnRH treatment in intestine, gill, heart, stomach, kidney and spleen. This study shows that GnRH can exert direct effects in both reproductive and non-reproductive somatic tissues of the gilthead seabream.     

Conservation of genomic imprinting at the NDN, MAGEL2 and MEST loci in pigs

Imprinted genes have important effects on the regulation of fetal growth, development, and postnatal behavior. However, the study of imprinted genes has been limited in mammalian species other than human and mouse. Therefore, the study of porcine imprinted genes is useful for defining the extent of conservation of genomic imprinting among different species. In this study, the imprinting status of porcine NDN, MAGEL2 and MEST genes was determined by direct sequencing of the cDNAs and detection of single nucleotide polymorphisms (SNPs) identified in individuals from reciprocal crosses between Meishan and Large White pigs for allele discrimination. The analysis was carried out in 13 different tissues (skeletal muscle, fat, pituitary gland, heart, lung, liver, kidney, spleen, stomach, small intestine, uterus, ovary and testis) from 12 two-month-old piglets. Imprinting analysis showed that NDN and MAGEL2 were paternally expressed in all tissues where the genes were expressed as in human and mouse. Interestingly, MEST showed tissue-specific imprinting, being paternally expressed in skeletal muscle, fat, pituitary gland, heart, kidney, lung, stomach and uterus, and maternally expressed in spleen and liver.     

Identification of two insulin-like growth factor IIs in the Japanese eel, Anguilla japonica: cloning, tissue distribution, and expression after growth hormone treatment and seawater acclimation

To better understand the role of IGFs in Japanese eel, Anguilla japonica, we cloned insulin-like growth factor-II (IGF-II) cDNAs and examined their mRNA expression in several tissues. Two eel IGF-II cDNAs, eIGF-II-1 and eIGF-II-2, were cloned from the liver. A signal peptide and a mature peptide of both preproIGF-IIs were composed of 47 amino acids (aa) and 69 aa, but they differed at 17 aa and 13 aa, respectively. The E domain of eIGF-II-1 was 49 aa longer than that of eIGF-II-2, and differed at 22 aa within 52 aa. The highest eIGF-II-1 and II-2 mRNA levels were observed in the liver, with detectable levels also found in all tissues examined. The eIGF-II-1 mRNA levels in the liver, heart, and muscle were higher in females than in males, whereas those in the stomach and intestine were lower in the females. The eIGF-II-2 mRNA levels in the liver and swim-bladder were also higher in females than in males whereas those in the stomach, spleen, and intestine were lower in the females. The eIGF-II-1 mRNA levels in the liver were higher in large compared to small glass eels, while the eIGF-II-2 mRNA levels did not correlate with body weight. Both eIGF-II mRNA levels in the liver increased after eel GH treatments in vivo and in vitro. No differences in both eIGF-II mRNA levels were observed in the gills, liver, stomach and whole kidney between seawater- and freshwater-reared eels.